Plantibody purification is not as efficient as antibody purification from serum, ascites, or mammalian cell cultures. It is characterized by the application of inefficient plantibody solid-liquid extraction systems, low plantibody recovery, and short lifetimes of expensive chromatography matrices. To overcome it, several protocols of liquid-liquid aqueous two-phase extraction (ATPE) combined with affinity chromatography were previously studied to purify the CB.Hep-1 monoclonal antibody, which showed an unexpectedly high recovery. However, a study of ATPE combined with several affinity chromatography matrices to purify plantibodies has not been reported so far. Therefore, a combination of the best ATPE protocol with five specific affinity chromatography matrices to purify a plantibody for vaccine manufacturing is described in this study. Positive outcomes from plantibody recovery (%), specific activity (%), yield (mg purified IgG/L of leaf extract), and productivity (mg purified IgG/L of leaf extract/h) were achieved. Plantibody purity did not show statistical differences among all samples (> 97%, p < 0.05), and protein A leakage was thousands of times smaller than toxic protein A for non-human primates. In summary, the combination of ATPE (10% PEG 4000/15% K2PO4, pH 5.5) with two specific affinity resins were well-suited for large-scale plantibody purification from tobacco plant leaves...
Tag: <span>plantibody</span>
Regulatory agencies routinely announce changes in the GMP (Good Manufacturing Practices) norms and quality standards designed to guarantee that every product retains characteristics required for its pharmaceutical use.The alignment with emerging regulatory requirements is mandatory, therefore, to assure production process consistency, as well as the safety and efficacy of the finished product…
Despite the existence of effective vaccines against Hepatitis B virus, the infection with it remains an important problem worldwide due to its association with hepatocellular carcinoma. Several procedures have been used to purify the Hepatitis B surface antigen (HBsAg) for immunization purposes. Immuno-purification using HBsAg-specific murine monoclonal antibodies (MAbs) has been one of the most successful strategies for such a purpose due to the high antigen selectivity (high affinity) of MAbs…
Regulatory agencies such as the FDA require the structure and amino acid sequence characterization of recombinant monoclonal antibodies (MAbs) to grant marketing approval. Characterizing such complex, inherently heterogeneous molecules is a significant analytical challenge that requires a broad array of physicochemical tests. Mass spectrometry (MS) is an essential tool for characterizing protein identity, functions, substrate specificity and amino acid sequence (AAS) of recombinant MAb biotherapeutics as it complements, or in some cases supersedes the utility of traditional biological methods. For some of the most important proteomic applications, the high sensitivity and accuracy provided by modern MS has allowed the unequivocal protein characterization…
Proteins are widely used in research, medicine and industry, but its extraction from their natural sources can be difficult, tedious and expensive. Therefore, a simple and inexpensive system that allows large-scale production of safe recombinant proteins will always be highly desirable. Traditional production systems that use microbial, insect and mammalian cell cultures have drawbacks, in terms of cost, scalability and product safety. Several studies have shown that molecular farming in plants has many practical, economic, and safety advantages as compared to these conventional methods. Thus, the use of plants for recombinant protein synthesis is gaining wide acceptance…